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Hypotaurine Supplementation Benefits in Cryopreservation (HYPOTAURINE)

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ClinicalTrials.gov Identifier: NCT04011813
Recruitment Status : Completed
First Posted : July 9, 2019
Last Update Posted : July 9, 2019
Sponsor:
Collaborators:
Université d'Auvergne, Institut National de la Santéet de la Recherche Médicale
Université d'Auvergne, Centre National de la Recherche Scientifique
University Hospital, Clermont-Ferrand, Hematology laboratory
Information provided by (Responsible Party):
University Hospital, Clermont-Ferrand

Brief Summary:
Although it is widely used, slow freezing can induce strong functional and nuclear spermatic alterations reducing the chances of pregnancy. The study objective is to determinate the effects of the combination of hypotaurine supplementation and spermatozoa selection by Density Gradient Centrifugation (DGC) on human sperm functions and DNA quality during a freezing-thawing cycle.

Condition or disease Intervention/treatment
Cryopreservation Human Spermatozoa Parameters Hypotaurine Supplementation Spermatozoa DNA Alterations Other: Hypotaurine : antioxidant and osmoregulator

Detailed Description:
This prospective study was performed on surplus semen after a density gradient centrifugation-frozen-thawing cycle. Samples were obtained from men undergoing routine semen analysis at the Center for Reproductive Medicine. Spermatozoa were selected by density gradient centrifugation, washed and frozen using a programmable device. Each step was performed in parallel with (H+ arm) or without (H- arm) 50mM hypotaurine supplementation. After thawing, investigator team compared for both conditions the total and progressive mobility, vitality, integrity of the acrosome, markers of Protein Kinase A (PKA) dependent capacitation intracellular signaling pathway and nuclear quality by measuring chromatin packaging, DNA fragmentation and oxidation and vacuoles presence in the spermatozoa head.

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Study Type : Observational
Actual Enrollment : 33 participants
Observational Model: Cohort
Time Perspective: Prospective
Official Title: Official Title: Hypotaurine Supplementation in Freezing and Preparation Media Improves Human Sperm DeoxyriboNucleic Acid (DNA) and Fertilizing Ability
Actual Study Start Date : May 1, 2014
Actual Primary Completion Date : August 1, 2018
Actual Study Completion Date : May 1, 2019

Group/Cohort Intervention/treatment
Control arm, "H-"
semen treated without hypotaurine supplementation in density gradient centrifugation, washing and cryopreservation media
Experimental arm, "H+"
semen treated with a 50mM hypotaurine supplementation in density gradient centrifugation, washing and cryopreservation media
Other: Hypotaurine : antioxidant and osmoregulator

Hypotaurine has protective effects on sperm motility, capacitation and acrosome reaction and reduces apoptotic markers.

Hypotaurine (50mM) was added in density gradient centrifugation, washing and cryopreservation media washing and cryopreservation media before spermatozoa freezing





Primary Outcome Measures :
  1. chromatin packaging labelled using aniline blue and chromomycin A3 [ Time Frame: Day 0 ]
    Comparison after a cycle of freezing-thawing for both conditions, with or without hypotaurine supplementation of chromatin packaging labelled using aniline blue and chromomycin A3

  2. DNA fragmentation using TUNEL assay [ Time Frame: Day 0 ]
    Comparison after a cycle of freezing-thawing for both conditions, with or without hypotaurine supplementation of DNA fragmentation using TUNEL assay

  3. DNA oxidation assessed by 8-OHdG immunodetections [ Time Frame: Day 0 ]
    Comparison after a cycle of freezing-thawing for both conditions, with or without hypotaurine supplementation of DNA oxidation assessed by 8-OHdG immunodetections

  4. vacuoles presence in the spermatozoa head using Motile Sperm Organelle Morphology Examination (MSOME) [ Time Frame: Day 0 ]
    Comparison after a cycle of freezing-thawing for both conditions, with or without hypotaurine supplementation of vacuoles presence in the spermatozoa head using Motile Sperm Organelle Morphology Examination (MSOME)


Secondary Outcome Measures :
  1. vitality using Eosin Nigrosin [ Time Frame: Day 0 ]
    Comparison after a cycle of freezing-thawing for both conditions, with or without hypotaurine supplementation of vitality using Eosin Nigrosin

  2. motility total and progressive [ Time Frame: Day 0 ]
    Comparison after a cycle of freezing-thawing for both conditions, with or without hypotaurine supplementation of motility total and progressive

  3. integrity of the acrosome using Fluorescein IsoThioCyanate-Pisum Sativum Agglutinin (FITC-PSA) labelling [ Time Frame: Day 0 ]
    Comparison after a cycle of freezing-thawing for both conditions, with or without hypotaurine supplementation of integrity of the acrosome using Fluorescein IsoThioCyanate-Pisum Sativum Agglutinin (FITC-PSA) labelling

  4. markers of PKA-dependent capacitation intracellular signaling pathway assessing western blot [ Time Frame: Day 0 ]
    Comparison after a cycle of freezing-thawing for both conditions, with or without hypotaurine supplementation of markers of PKA-dependent capacitation intracellular signaling pathway assessing western blot



Information from the National Library of Medicine

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Ages Eligible for Study:   18 Years and older   (Adult, Older Adult)
Sexes Eligible for Study:   Male
Accepts Healthy Volunteers:   No
Study Population
Men undergoing routine semen analysis at the Center for Reproductive Medicine
Criteria

Inclusion Criteria:

  • None

Exclusion Criteria:

  • None

Information from the National Library of Medicine

To learn more about this study, you or your doctor may contact the study research staff using the contact information provided by the sponsor.

Please refer to this study by its ClinicalTrials.gov identifier (NCT number): NCT04011813


Sponsors and Collaborators
University Hospital, Clermont-Ferrand
Université d'Auvergne, Institut National de la Santéet de la Recherche Médicale
Université d'Auvergne, Centre National de la Recherche Scientifique
University Hospital, Clermont-Ferrand, Hematology laboratory
Investigators
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Principal Investigator: Florence BRUGNON, MD, PhD University Hospital, Clermont-Ferrand
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Responsible Party: University Hospital, Clermont-Ferrand
ClinicalTrials.gov Identifier: NCT04011813    
Other Study ID Numbers: 2014 HYPOTAURINE
First Posted: July 9, 2019    Key Record Dates
Last Update Posted: July 9, 2019
Last Verified: June 2019

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Studies a U.S. FDA-regulated Drug Product: No
Studies a U.S. FDA-regulated Device Product: No
Keywords provided by University Hospital, Clermont-Ferrand:
cryopreservation
density gradient centrifugation
DNA fragmentation, oxidation, chromatin packaging
antioxidant hypotaurine supplementation
cryo-capacitation
human spermatozoa fertilizing ability
Additional relevant MeSH terms:
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Antioxidants
Hypotaurine
Molecular Mechanisms of Pharmacological Action
Protective Agents
Physiological Effects of Drugs